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Western Blot Analysis:
Untreated (lanes 1, 3, 5) and Calyculin A/Okadaic Acid-treated (lanes 2, 4, 6) IR/IRS1 transfected CHO cell lysate was res...
Western Blot Analysis:
Untreated (lanes 1) and Insulin-treated (lanes 2) HEK293 cell lysate was resolved by electrophoresis, transferred to PVDF ...
Anti-phospho-IRS1 (Ser302 mouse/ Ser307 human), clone 8.1.2
| Species Reactivity | Key Applications | Host | Format | Antibody Type |
|---|---|---|---|---|
| H, M, R, Mk, B, Ca, Po | IH, IF, IP, WB | Mouse | Purified | Monoclonal Antibody |
Description:
Anti-phospho-IRS1 (Ser302 mouse/ Ser307 human), clone 8.1.2
Specificity:
This antibody recognizes IRS1 phosphorylated at Ser302 (mouse) and Ser307 (human).
Molecular Weight:
~185 kDa.
Epitope:
Phosphorylated Ser302
Immunogen:
Synthetic linear peptide corresponding to amino acids at and around phosphorylated Ser302 of mouse IRS1.
Modifications:
Phosphorylation
Clone:
8.1.2
Isotype:
IgG1κ
Background Information:
IRS1 (Insulin Receptor Substrate 1) transmits insulin signals via metabolic and mitogenic pathways. IRS1 is heavily phosphorylated on both serine and tyrosine residues. These phosphorylated tyrosines enable IRS to act as a docking protein that binds SH2 domains of such proteins as PI3 Kinase (phosphatidylinositol 3-kinase) and GRB2, resulting in activation. Over expression and phosphorylation of serine is associated with insulin resistance and breast cancer. Some of the more notable phosphorylation sites are Ser302 that is phosphorylated following insulin stimulation. Ser307, phosphorylated by JNK and IKK, is a key regulatory site that appears to disrupt the IRS1/IR interaction and inhibits insulin-mediated activation of the PI3 kinase and MAPK pathways, and Ser636/639 that is known to be phosphorylated by p70S6K downstream of mTOR and acts as a negative feedback loop.
Species Reactivity:
- Human
- Mouse
- Rat
- Monkey
- Bovine
- Canine
- Pig
Species Reactivity Note:
Human, Mouse, and Rat. Predicted to cross-react with many other species based on 100% sequence homology with immunogen.
Application Notes:
Immunocytochemistry:
IR/IRS transfected CHO cells were stained with a 1:500 dilution of the antibody. A seconday anti-mouse Cy3 conjugate was used for visualizaion.
Immunoprecipition:
This antibody was used at a 1:1,000 to immunoprecipitate IRS1 from 5L/mL IR/IRS1 transfected CHO cells (performed by an independent lab).
IR/IRS transfected CHO cells were stained with a 1:500 dilution of the antibody. A seconday anti-mouse Cy3 conjugate was used for visualizaion.
Immunoprecipition:
This antibody was used at a 1:1,000 to immunoprecipitate IRS1 from 5L/mL IR/IRS1 transfected CHO cells (performed by an independent lab).
Quality Assurance:
Evaluated by western blot on IRS1 transfected CHO +/- Insulin lysate.
Western Blot Analysis:
1:1,000 dilution of this antibody was used to detect IRS1 phosphorylated on Ser302 in IRS/IR transfected CHO -/+ Calyculin A/ Okadaic Acid-treated cell lysate.
Western Blot Analysis:
1:1,000 dilution of this antibody was used to detect IRS1 phosphorylated on Ser302 in IRS/IR transfected CHO -/+ Calyculin A/ Okadaic Acid-treated cell lysate.
Purification Method:
Protein G purfied
Presentation:
Purified mouse monoclonal antibody in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide.
Storage Conditions:
Stable for 1 year at 2-8ºC from date of receipt.
UniProt Number:
Entrez Gene Number:
Gene Symbol:
- HIRS-1
- IRS-1
Alternate Names:
insulin receptor substrate 1
Usage Statement:
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
Key Applications:
- Immunohistochemistry
- Immunofluorescence
- Immunoprecipitation
- Western Blotting
UniProt Summary:
FUNCTION: May mediate the control of various cellular processes by insulin. When phosphorylated by the insulin receptor binds specifically to various cellular proteins containing SH2 domains such as phosphatidylinositol 3-kinase p85 subunit or GRB2. Activates phosphatidylinositol 3-kinase when bound to the regulatory p85 subunit By similarity.
SUBUNIT STRUCTURE: Interacts with the NPXY motif of tyrosine-phosphorylated IGF1R and INSR via the PTB domain. Binds to phosphatidylinositol 3-kinase p85 subunit via the phosphorylated YXXM motifs. Binds ROCK1. Binds to UBTF and PIK3CA in nuclear extracts By similarity. Interacts with SOCS7.
PTM: Serine phosphorylation of IRS1 is a mechanism for insulin resistance. Ser-312 phosphorylation inhibits insulin action through disruption of IRS1 interaction with the insulin receptor By similarity.
Phosphorylation of Tyr-896 is required for GRB2-binding By similarity.
Polymorphism The Arg-971 polymorphism impairs the ability of insulin to stimulate glucose transport, glucose transporter translocation, and glycogen synthesis by affecting the PI3K/AKT1/GSK3 signaling pathway. The polymorphism at Arg-971 may contribute to the in vivo insulin resistance observed in carriers of this variant. Arg-971 could contribute to the risk for atherosclerotic cardiovascular diseases associated with non-insulin-dependent diabetes mellitus (NIDDM) by producing a cluster of insulin resistance-related metabolic abnormalities. In insulin-stimulated human endothelial cells from carriers of the Arg-971 polymorphism, genetic impairment of the IRS1/PI3K/PDPK1/AKT1 insulin signaling cascade results in impaired insulin-stimulated nitric oxide (NO) release and suggested that this may be a mechanism through which the Arg-971 polymorphism contributes to the genetic predisposition to develop endothelial dysfunction and cardiovascular disease. The Arg-971 polymorphism not only reduces phosphorylation of the substrate but allows IRS1 to act as an inhibitor of PI3K, producing global insulin resistance.
DISEASE: Polymorphisms in IRS1 may be involved in the etiology of non-insulin-dependent diabetes mellitus (NIDDM) [MIM:125853]. Ref.18 Ref.19 Ref.21 Ref.24 Ref.25
SIMILARITIY: Contains 1 IRS-type PTB domain.
Contains 1 PH domain.
SUBUNIT STRUCTURE: Interacts with the NPXY motif of tyrosine-phosphorylated IGF1R and INSR via the PTB domain. Binds to phosphatidylinositol 3-kinase p85 subunit via the phosphorylated YXXM motifs. Binds ROCK1. Binds to UBTF and PIK3CA in nuclear extracts By similarity. Interacts with SOCS7.
PTM: Serine phosphorylation of IRS1 is a mechanism for insulin resistance. Ser-312 phosphorylation inhibits insulin action through disruption of IRS1 interaction with the insulin receptor By similarity.
Phosphorylation of Tyr-896 is required for GRB2-binding By similarity.
Polymorphism The Arg-971 polymorphism impairs the ability of insulin to stimulate glucose transport, glucose transporter translocation, and glycogen synthesis by affecting the PI3K/AKT1/GSK3 signaling pathway. The polymorphism at Arg-971 may contribute to the in vivo insulin resistance observed in carriers of this variant. Arg-971 could contribute to the risk for atherosclerotic cardiovascular diseases associated with non-insulin-dependent diabetes mellitus (NIDDM) by producing a cluster of insulin resistance-related metabolic abnormalities. In insulin-stimulated human endothelial cells from carriers of the Arg-971 polymorphism, genetic impairment of the IRS1/PI3K/PDPK1/AKT1 insulin signaling cascade results in impaired insulin-stimulated nitric oxide (NO) release and suggested that this may be a mechanism through which the Arg-971 polymorphism contributes to the genetic predisposition to develop endothelial dysfunction and cardiovascular disease. The Arg-971 polymorphism not only reduces phosphorylation of the substrate but allows IRS1 to act as an inhibitor of PI3K, producing global insulin resistance.
DISEASE: Polymorphisms in IRS1 may be involved in the etiology of non-insulin-dependent diabetes mellitus (NIDDM) [MIM:125853]. Ref.18 Ref.19 Ref.21 Ref.24 Ref.25
SIMILARITIY: Contains 1 IRS-type PTB domain.
Contains 1 PH domain.
Qty/Pk:
100 µg
Format:
Purified
Host:
Mouse
Product Name:
Anti-phospho-IRS1 (Ser302 mouse/ Ser307 human), clone 8.1.2
Concentration:
1 mg/mL
Antibody Type:
Monoclonal Antibody